Статті в журналах з теми "Purification par extraction liquide"

Polycyclic aromatic hydrocarbons (PAHs) are among the organic pollutants sought both in the various environmental compartments (water, soil, air, etc.) and in food intended for human consumption. They are a group of persistent organic pollutants that are potentially mutagenic and carcinogenic. Thus, the present study aims to determine the PAH levels in the flesh of local poultry also called bicycle chickens (gallus gallusdomesticus). Ten (10) composite samples of chickens were collected in Karakoro, a locality located in the department of Korhogo in the north of Cote dIvoire. After the extraction and purification steps, the samples were analyzed on a Shimadzu brand HighPerformance Liquid Chromatograph (HPLC) equipped with a fluorimetric detector. The measured PAHs, eight (8) molecules in number, are classified among the priority substances to be researched in the context of food safety (fluoranthene, pyrene, benzo(k)fluoranthene, benzo(a)pyrene, indeno(1 ,2,3-cd) pyrene, benzo (g,h,i) perylene, benzo (a) anthracene, benzo (b) fluoranthene). The results obtained showed the presence of these molecules in the samples analyzed at concentrations higher than the European Union (EU) standard for some. On the other hand, all the PAHs determined have concentrations lower than the Codex Alimentarius standard.

The liquid–liquid extraction of cobalt(II) from sulfate medium with N-(2- hydroxybenzylidene)aniline is studied with the following parameters: pH, concentration of the extractant and the nature of diluent. The solvent extraction of cobalt(II) in different diluents leads to third phase formation. The stoichiometry coefficients of the extracted complexes are determined by the slope analysis method. The extaction constants are evaluated for the different diluents. The extraction is better in the following order: cyclohexane > toluene > chloroform.

Résumé Au cours de ce travail, nous avons étudié l’extraction liquide - liquide et le transport du plomb à travers des membranes polymériques en utilisant le chlorure de tricapryle ammonium (Aliquat 336) comme transporteur. L’extraction du plomb par Aliquat 336 nous a permis de mettre au point les conditions optimales d’extraction et de déterminer les rendements d’extraction. Les expériences d’extraction liquide - liquide ont montré qu’un maximum de rendement était obtenu après huit minutes d’agitation à une vitesse de 2 400 (rpm). Les pourcentages d’extraction obtenus varient entre 80 à 94 % et donc une très bonne élimination du plomb a été réalisée (la concentration initiale du plomb varie entre 2 10‑6 M et 30 10‑6 M). Nous avons préparé par la suite des membranes à base du polymère triacétate de cellulose (TAC) plastifiées par le tris-ethyl-hexyl-phosphate (TEHP) et modifiées par l’extractant Aliquat 336 utilisé comme transporteur. Les membranes élaborées ont été également caractérisées par spectroscopie infrarouge à transformée de Fourier (FTIR). Les valeurs obtenues pour les épaisseurs (entre 10 et 20 µm) et les densités sont tout à fait comparables à celles des supports commerciaux utilisés pour la préparation des membranes liquides supportées. L’ajout du plastifiant a donné une très bonne hydrophobie des membranes élaborées. Les expériences du transfert du plomb à travers les nouvelles membranes ont montré que les flux augmentent considérablement avec la concentration du transporteur pour atteindre un maximum à partir de 10‑3 M. D’autres paramètres caractérisant le transport (concentration initiale du métal et le pH) ont été déterminés. Un bon rendement d’élimination du plomb a été obtenu dans la gamme de pH très acide (1 ≤ pH ≤ 2).

AbstractA high yielding synthesis of 1,3-oxazinan-2-ones starting from 3-amino-1-propanols and ethylene carbonate (EC) in the presence of catalytic amount of triazabicyclodecene (TBD) is herein reported. The formation of six-membered cyclic carbonates was achieved by intermolecular cyclization reaction via double BAc2 mechanism. Cyclization reactions have been carried out in neat as EC acted both as solvent and reagent. Pure 1,3-oxazinan-2-ones were isolated in high yield by simple liquid-liquid extraction. Further purification can be achieved by recrystallization. The reaction resulted of general application on different substrates including an aryl bis(3-amino-proan-1-ol) compound.

Suite à la détection de quantités importantes de composés perfluorés dans des prélèvements d’air, de sol, d’eau ainsi que de lait maternel, deux arrêtés préfectoraux de la préfecture du Rhône, datant de mai 2022, ont défini les limites de quantification de ces contaminants environnementaux à 10 ng/L dans les eaux usées. Ces composés très persistants et considérés comme des perturbateurs endocriniens peuvent avoir des effets non négligeables sur la santé humaine. La nécessité de développer de nouveaux outils de détection et de quantification de ces molécules dans les eaux de rejets est donc urgente et indispensable. Afin de répondre au besoin de quantifier ces contaminants environnementaux à de basses teneurs, une méthode d’analyse par chromatographie liquide couplée à la spectrométrie de masse en tandem, a été développée. Au préalable, les échantillons subissent une étape de préparation d’échantillons permettant d’extraire les composés d’intérêt des eaux usées par une extraction liquide-liquide. Cette méthode a été validée selon la norme NF T90-210 et permet la détection de 25 composés perfluorés et la quantification de 15 d’entre eux, jusqu’à des teneurs de l’ordre de 10 ng/L, comme demandé par les arrêtés préfectoraux. Les taux de recouvrement s’élèvent entre 87 et 117 % avec des coefficients de variation de fidélité intermédiaire compris entre 6 et 25 %. La méthode analytique est ainsi accréditée Cofrac pour 15 composés perfluorés. Enfin, cette dernière a été appliquée à plus d’une centaine d’échantillons d’eaux usées, dits réels, démontrant la pollution d’un grand nombre d’entre eux aux composés perfluorés. Parmi ces composés, trois sont les plus souvent retrouvés dans les eaux résiduaires, il s’agit du PFOA, du PFNA et du PFHxA.

Objectif : Contribuer au contrôle de qualité des moustiquaires imprégnées d’insecticides par la technique de la Chromatographie liquide à haute performance pour assurer leur efficacité dans la prévention du paludisme au Mali. Matériel et Méthodes : les échantillons étaient constitués de moustiquaires achetées ou obtenues gratuitement. L’échantillonnage a consisté à une coupure aléatoire de petits morceaux sur les cinq côtés de la moustiquaire puis une coupure d’un carré modèle de 10*10 cm. Au total quatre moustiquaires ont fait l’objet d’analyse. Le matériel utilisé pour l’analyse des moustiquaires était la Chromatographie liquide haute performance (HPLC) couplée à un Détecteur à Barrette de Diode. La méthode d’analyse était basée sur une extraction solide/liquide avec comme solvant d’extraction l’Acétone pendant une période de deux heures puis évaporer à sec l’extrait. L’extrait sec a été récupéré avec 1 mL d’Acétonitrile pour analyse. Résultats : Tous les échantillons étaient à base de polyester, sauf un qui était en polyéthylène et un poids variant entre 452,3 et 651,0 grammes. Les conditions chromatographiques utilisées ont permis d’identifier et doser la Deltaméthrine dans les échantillons de moustiquaires à un temps de rétention de 3,27 minutes. Les différentes gammes de concentrations étaient proportionnelles à la surface des pics obtenues. Un échantillon sur quatre était sous dosé donc non conforme. Conclusion : La méthode ainsi mise en place permettra le dosage de la Deltaméthrine dans les différents matériaux de moustiquaires afin d’assurer la surveillance de la qualité des moustiquaires au Mali.

<p style="text-align: justify;">Un certain nombre de composés phénoliques simples des vins peuvent être détectés et quantifiés par chromatographie liquide à haute performance sans extraction préalable. Les résultats enregistrés sur des vins jeunes montrent en particulier que lors des six premiers mois les teneurs en acides gallique et en catéchlne augmentent, alors que celle en tyrosol diminue. En ce qui concerne les acides hydroxycinnamoyl-tartriques, on observe souvent un enrichissement, mais ceci affecte peu les teneurs en acides hydroxycinnamiques totaux.</p><p style="text-align: justify;">Dans le cas des vins vieux on constate essentiellement la très haute teneur en tyrosol, particulièrement sur les plus anciens millésimes.</p>

Molecular characterization of the microbial populations of soils and sediments contaminated with polycyclic aromatic hydrocarbons (PAHs) is often a first step in assessing intrinsic biodegradation potential. However, soils are problematic for molecular analysis owing to the presence of organic matter, such as humic acids. Furthermore, the presence of contaminants, such as PAHs, can cause further challenges to DNA extraction, quantification, and amplification. The goal of our study was to compare the effectiveness of four commercial soil DNA extraction kits (UltraClean Soil DNA Isolation kit, PowerSoil DNA Isolation kit, PowerMax Soil DNA Isolation kit, and FastDNA SPIN kit) to extract pure, high-quality bacterial and eukaryotic DNA from PAH-contaminated soils. Six different contaminated soils were used to determine if there were any biases among the kits due to soil properties or level of contamination. Extracted DNA was used as a template for bacterial 16S rDNA and eukaryotic 18S rDNA amplifications, and PCR products were subsequently analyzed using denaturing gel gradient electrophoresis (DGGE). We found that the FastDNA SPIN kit provided significantly higher DNA yields for all soils; however, it also resulted in the highest levels of humic acid contamination. Soil texture and organic carbon content of the soil did not affect the DNA yield of any kit. Moreover, a liquid–liquid extraction of the DNA extracts found no residual PAHs, indicating that all kits were effective at removing contaminants in the extraction process. Although the PowerSoil DNA Isolation kit gave relatively low DNA yields, it provided the highest quality DNA based on successful amplification of both bacterial and eukaryotic DNA for all six soils. DGGE fingerprints among the kits were dramatically different for both bacterial and eukaryotic DNA. The PowerSoil DNA Isolation kit revealed multiple bands for each soil and provided the most consistent DGGE profiles among replicates for both bacterial and eukaryotic DNA.

Abstract A liquid chromatographic (LC) procedure for determining 10 biogenic amines in cheese is described. The method is based on ion-pair chromatography on a reversed-phase column with postcolumn derivatization with o-phthaldialdehyde and fluorometric detection. It allowed simultaneous determination of 10 amines in &lt;80 min: histamine, tyramine, tryptamine, 2-phenylethylamine, serotonin, agmatine,spermine, spermidine, putrescine, and cadaverine. Linearity for each amine was observed between 0.5 and 6.0 μg/mL. Detection limits ranged from 0.004 to 0.009 μg/20 μL, and determination limits ranged from 0.066 to 0.149 mg/100 g. Amino acids and other amines did not interfere with determination of biogenic amines. Three extractantsmethanol, hydrochloric acid, and trichloroacetic acidwere compared in their efficiency to recover aminesfrom spiked samples. Purification of the cheese extract was required prior to LC to avoid interference from compounds in the cheese matrix. Hydrochloric acid extraction followed by purification with diethyl ether gave best recoveries for all the amines (75.5-112.3%). The method is simple, fast, and reliable. It can be used to study the technological and toxicological implications of biogenic amines in cheeses.

The priority task of modern biotechnology is development of the rational technologies for the microbial synthesis of practically important products. Among these products, a significant place belongs to surfactants (biosurfactants), which are widely used in many sectors of the economy. The most problematic stage of the biosurfactants production is isolation from the post fermentative cultural liquid of bacteria-producers. Improving the efficiency of the biosurfactants production is highly dependent on rational approaches to the target products isolation. In this regard, there is an increasing need for rational, scientifically substantiated methods for their isolation and purification. Therefore, the aim of the presented work was to determine the optimal extractants for the isolation of rhamnolipid surfactants – metabolites of bacteria of Pseudomonas sp. PS-17 strain. For this purpose, the extraction process of rhamnolipids from the post fermentative cultural liquid supernatant has been investigated. The optimal extractants were selected among 13 organic solvents of different nature. Processing of the obtained experimental data by the method of multi-parameter equations of linearity of free energies (modified Koppel-Palm equation) made it possible to establish the relationship between the physicochemical properties of the extractants and amounts of the biosurfactants which were isolated from cultural liquid supernatant. It was shown that the data on the rhamnolipids extraction are adequately associated with the physicochemical characteristics of the solvents using a six-parameter linear equation. It was determined that the polarizability and molar volume are the main properties of solvents that affect the extraction process. The best extractants for the rhamnolipids isolation from cultural liquid supernatant of the Pseudomonas sp. PS-17 strain are the ethers. It can be explained by the presence of a lone pair of the electrons of oxygen in its molecule. The obtained results of the study are of scientific interest for isolation of the important and perspective biotechnological products – surface-active substances.

The introduction of synthetic dyes completely changed the industrial production and use of colorants for art materials. From the synthesis of the first synthetic dye, mauveine, in 1856 until today, artists have enjoyed a wider range of colors and selection of chemical properties than was ever available before. However, the introduction of synthetic dyes introduced a wider variety and increased the complexity of the chemical structures of marketed dyes. This work looks towards the analysis of synthetically dyed objects in heritage collections, applying an extraction protocol based on the use of ammonia, which is considered favorable for natural anthraquinone dyes but has never before been applied to acid synthetic dyes. This work also presents an innovative cleanup step based on the use of an ion pair dispersive liquid–liquid microextraction for the purification and preconcentration of historical synthetic dyes before analysis. This approach was adapted from food science analysis and is applied to synthetic dyes in heritage science for the first time in this paper. The results showed adequate recovery of analytes and allowed for the ammonia-based extraction method to be applied successfully to 15 samples of suspected azo dyes from the Azienda Coloranti Nazionali e Affini (ACNA) synthetic dye collection, identified through untargeted HPLC-HRMS analyses.

Operating in a cell of water/CHCl3/water, it is shown that there is a preferential transport of Ca2+/Mg2+ by A.23187. On the basis of measurements of flux of extraction and of flux of liberation in a half-cell of water/CHCl3, it is shown that the selectivity of the transport is related to the kinetics of liberation, which is markedly different for the two ions and which could be explained by the structure of the complex dimers that are formed, according to the corresponding crystallographic studies. The methylation of the —NHCH3 entails an inversion of the selectivity in the transport

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous environmental and processing contaminants generated by both spontaneous and anthropogenic incomplete combustion processes of organic matter. Contamination of PAHs in vegetable oils can result from several factors and processes, including environmental contamination, oil processing, and migration from food contact materials. The determination of PAHs in edible oil presents a challenge because of the complexity of the matrix. Since PAHs are present at lower levels than triglycerides, it is necessary to isolate the compounds of interest from the rest of the matrix. To this purpose, a new purification approach based on a double solid-phase extraction (SPE) step followed by high performance liquid chromatography–fluorometric detector (HPLC-FLD) analysis was developed. The method involves a first purification step by using a 5 g silica SPE cartridge, previously washed with dichloromethane (20 mL), dried completely, and then conditioned with n-hexane (20 mL). The triglycerides are retained by the silica, while the PAH-containing fraction is eluted with a mixture of n-hexane/dichloromethane (70/30, v/v). After evaporation, the residue is loaded on a 5 g amino SPE cartridge and eluted with n-hexane/toluene (70/30, v/v) before HPLC-FLD analysis. The focus was the evaluation of the contribution of the various phases of the pomace oil supply chain in terms of the heavy PAHs (PAH8) concentration. Data collected showed that pomace contamination increased (by 15 times) as storage time increased. In addition, the process of pomace drying, which is necessary to reduce its moisture content before solvent extraction of the residual oil, appeared to significantly contribute to the total heavy PAHs content, with increases in value by up to 75 times.

<p style="text-align: justify;">Les différentes techniques d'extraction et de séparation des constituants de l'arôme des vins sont décrites et leurs avantages, inconvénients et domaines d'application mis en évidence.</p><p style="text-align: justify;">Les techniques d'extraction mettant en jeu un solvant en phase liquide ou supercritique, une distillation ou une concentration de l'espace de tête sont successivement étudiées. En ce qui concerne les séparations des constituants des extraits, l'accent est mis sur le type de phases à utiliser en CPV et sur les techniques de préfractionnement des extraits aromatiques par HPLC et chromatographie en deux dimensions.</p><p style="text-align: justify;">La difficulté d'obtenir un extrait olfactivement représentatif de l'arôme du vin est mis en évidence par l'analyse sensorielle.</p><p style="text-align: justify;">+++</p><p style="text-align: justify;">The different extraction and separation techniques used for studying volatile wine constituents are described with their advantages, disadvantages and specificity of application.</p><p style="text-align: justify;">Different extraction techniques involving a solvent in the liquid or supercritical phase, a distillation step or a concentration of the head-space are successively reviewed. Concerning separation of volatile constituents, some emphasis was given to the type of GC phase which an the most suitable and to different HPLC or two dimensional GC techniques which can be used to get more simple fractions from the crude extract.</p><p style="text-align: justify;">The author outlines from result of sensory analysis the importance and difficulty to obtain an extract the odour of which is similar to that of the wine.</p>

Les médicaments à usage humain ou vétérinaire se retrouvent, par différents moyens, dans les milieux aquatiques et notamment dans les filières de potabilisation des eaux. Les médicaments sont en partie métabolisés après ingestion (le pourcentage dépend du principe actif et est compris entre 1 et 99 %). Des médicaments se retrouvent alors dans les eaux destinées à la consommation humaine avec des concentrations allant du ng/L à la centaine de ng/L. Différents types de métabolites existent, et parmi eux les dérivés conjugués. Ils peuvent représenter 90 % des métabolites d’un principe actif. Le travail présenté ici décrit le développement d’une méthode analytique par « Solid Phase Extraction » (SPE) en ligne – chromatographie liquide haute performance – spectrométrie de masse en tandem (LC-MS/MS) pour la détermination et la quantification de principes actifs et leurs dérivés conjugués dans les eaux de surface et en cours de potabilisation. Ces dérivés conjugués sont susceptibles de subir une déconjugaison lors des traitements de potabilisation conduisant au relargage du principe actif. C’est pourquoi il est nécessaire de s’intéresser à ces molécules et en particulier les conjugués glucuronides. Comme tous les standards ne se sont pas disponibles dans le commerce, une nouvelle voie de synthèse en milieu aqueux a été explorée. Des analyses sont effectuées en entrée et sortie de filière de potabilisation d’eau afin de déterminer les concentrations présentes en principes actifs et leurs dérivés conjugués. Les limites de détection et de quantification obtenues sont de l’ordre du ng/L ou la dizaine de ng/L.

Kelor plant (Moringa oleifera Lamk) is a plant that grows naturally in a tropical climate. During this decoction of pirdot leaves plants are used by the people as antimicroba. The purpose of this study was to isolate triterpenoid compound from ethyl acetate extract of kelor leaves. The sample extraction was performed by maceration method using methanol as a solvent. Fractionation used n-hexana and etil asetat as a solvent. The separation and purification of the compound was carried out by column chromatography method followed by stain pattern analysis with thin layer chromatography (TLC). Characterization and elucidation of pure compound structures using UV, IR, 1H-NMR, 12C-NMR, HSQC, HMBC, and 1H-1H COSY, and compared with various literatures. Based on the result of structural elusidation, the pure isolate obtained is a pentacyclic triterpenoid group compound with the molecular formula C30H50O and the name of 3-hydroxy, 20 (29) -en, lupenol. The triterpenoid compound of 3-hidroxy, 20 (29) -en, lupenol is the first isolated and reported from leaves of kelor plant.

Kelor plant (Moringa oleifera Lamk) is a plant that grows naturally in a tropical climate. During this decoction of pirdot leaves plants are used by the people as antimicroba. The purpose of this study was to isolate triterpenoid compound from ethyl acetate extract of kelor leaves. The sample extraction was performed by maceration method using methanol as a solvent. Fractionation used n-hexana and etil asetat as a solvent. The separation and purification of the compound was carried out by column chromatography method followed by stain pattern analysis with thin layer chromatography (TLC). Characterization and elucidation of pure compound structures using UV, IR, 1H-NMR, 12C-NMR, HSQC, HMBC, and 1H-1H COSY, and compared with various literatures. Based on the result of structural elusidation, the pure isolate obtained is a pentacyclic triterpenoid group compound with the molecular formula C30H50O and the name of 3-hydroxy, 20 (29) -en, lupenol. The triterpenoid compound of 3-hidroxy, 20 (29) -en, lupenol is the first isolated and reported from leaves of kelor plant.

L’écorce de résineux, cultivés dans un but commercial, est une source potentielle précieuse de métabolites secondaires tels que les polyphénols. Les tannins, qui font partie des polyphénols présents dans l’écorce, sont utilisés dans la fabrication d’adhésifs et de résines, mais également en tant qu’agent de tannage, antibactérien, antifongique, antitermite et antioxydant. Peu d’informations existent à propos du rendement et de la composition des extraits d’écorce en fonction de la hauteur de l’échantillon prélevé dans le tronc ainsi qu’en fonction de la présence ou l’absence de branches. Cette étude a pour but d’examiner la variabilité des métabolites secondaires présents dans l’écorce d’Abies alba à la fois en fonction de la hauteur de l’échantillon prélevé dans un arbre, mais également la variabilité présente à des hauteurs spécifiques entre plusieurs arbres. La finalité de cette étude est de déterminer quelle fraction d’écorce contient le plus d’extractibles chez cette essence. Pour cela, huit arbres ont été sélectionnés dans lesquels un maximum de treize disques a été coupé tout le long du tronc. Ces échantillons ont été prélevés en bas du tronc à une hauteur de 30 cm du sol puis à différentes hauteurs Ces différentes hauteurs ont été choisies pour des raisons industrielles (hauteur limite pour le bois d’œuvre ou pour l’utilisation industrielle) mais également pour des raisons physiologiques (hauteur à la base du houppier, hauteur de la première branche verte…). Les échantillons prélevés ont été broyés puis extraits avec un mélange eau/éthanol (1 :1, v/v) en réalisant une extraction accélérée à chaud et sous pression. Une première étude quantitative est réalisée pour connaître la quantité d’extractibles totale présente dans l’écorce. La seconde étude est qualitative, afin de connaître quels types d’extractibles sont présents dans ces écorces. Ces extraits ont donc été examinés par chromatographie liquide couplée à un spectromètre UV-visible et un spectromètre de masse .Les résultats ont montré que la composition de l’extrait total d’écorce augmente en même temps que la hauteur dans le tronc. La proportion la plus élevée en composés polyphénoliques se trouve dans la section inférieure sous la couronne.

La domestication de Lippia multiflora, plantes aux multiples vertus en pharmacopée et médecine est un enjeu de taille en Côte d’Ivoire. Cependant, l’insuffisance de semences, due au faible taux de germination des graines, limite l’extension de sa culture. La présente étude avait pour objectif l’extraction, la purification et la caractérisation morpho-physiologique des graines de Lippia multiflora Moldenke (Verbenaceae). Il s’agissait plus spécifiquement de déterminer la pureté spécifique, le nombre de graines par unité de masse, le diamètre moyen et le taux d’humidité ; le taux de germination des graines de L. multiflora. Aussi, il s’est agi d’étudier l’impact du milieu sur le pouvoir germinatif des graines et de faire un suivi post-germination des plants en milieu réel. Après extraction des graines, des mesures physiques et des tests de germination ont permis de déterminer les caractéristiques morpho-physiologiques de celles-ci. L’étude a montré que le lot de graine étudié a une pureté spécifique de 70%, les graines ont un diamètre moyen de 0,34± 0.1 mm, une teneur en eau de 14±5.4 % et un taux de germination de 42,25%. Les tests de germination après un séjour prolongé dans divers milieux, révèlent qu’un milieu réfrigéré (7° C) confère une plus longue viabilité aux graines. Le suivi post-germination au champ montre une évolution régulière de la hauteur des plants, passant en moyenne de 2,58 cm à 8,8 cm au bout de 3 mois. Sur la même période, le nombre moyen de feuille varie de 4,03 à 21.Mots clés : Lippia multiflora, graine, caractérisation, germination, suivi post-germination. English Title: Morpho-physiological qualities and evaluation of the germination behavior of seeds of the savannah tea tree (Lippia multiflora Moldenke)The domestication of Lippia multiflora, plants with multiple virtues in pharmacopoeia and medicine is a major challenge in Côte d'Ivoire. However, the lack of seed, due to the low germination rate, limits the extension of its cultivation. The purpose of this study was the extraction, purification and morpho-physiological characterization of the seeds of Lippia multiflora Moldenke (Verbenaceae). More specifically, it involved determining the specific purity, the number of seeds per unit mass, the average diameter and the humidity rate; the germination rate of L. multiflora seeds. Also, it was a question of studying the impact of the environment on the germination power of seeds and of making a post-germination follow-up of the plants in real environment. After the seeds extraction, physical measurements and germination tests were carried out to determine the morpho-physiological characteristics. Results showed that the seed lot studied has a specific purity of 70%, an average diameter of 0.34 0.1 mm, a water content of 14 5.4% and a germination rate of 42.25%. Germination tests after a prolonged stay in various environments reveal that a refrigerated area (7 °C) confers a longer viability to the seeds. Post-germination monitoring in the field shows a steady increase in plant height from an average of 2.58 cm to 8.8 cm after 3 months. Over the same period, the average number of sheets varies from 4.03 to 21.Keywords: Lippia multiflora, seed, characterization, germination, post-germination monitoring.

AbstractNitrosylation of sulfhydryl (SH) groups of cysteine (Cys) moieties is an important post-translational modification (PTM), often on a par with phosphorylation. S-Nitrosoalbumin (ALB-Cys34SNO; SNALB) in plasma and S-nitrosohemoglobin (Hb-Cysβ93SNO; HbSNO) in red blood cells are considered the most abundant high-molecular-mass pools of nitric oxide (NO) bioactivity in the human circulation. SNALB per se is not an NO donor. Yet, it acts as a vasodilator and an inhibitor of platelet aggregation. SNALB can be formed by nitrosation of the sole reduced Cys group of albumin (Cys34) by nitrosating species such as nitrous acid (HONO) and nitrous anhydride (N2O3), two unstable intermediates of NO autoxidation. SNALB can also be formed by the transfer (S-transnitrosylation) of the nitrosyl group (NO+) of a low-molecular-mass (LMM) S-nitrosothiol (RSNO) to ALB-Cys34SH. In the present study, the effects of LMM thiols on the inhibitory potential of ALB-Cys34SNO on human washed platelets were investigated. ALB-Cys34SNO was prepared by reacting n-butylnitrite with albumin after selective extraction from plasma of a healthy donor on HiTrapBlue Sepharose cartridges. ALB-Cys34SNO was used in platelet aggregation measurements after extended purification on HiTrapBlue Sepharose and enrichment by ultrafiltration (cutoff, 20 kDa). All tested LMM cysteinyl thiols (R-CysSH) including l-cysteine and L-homocysteine (at 10 µM) were found to mediate the collagen-induced (1 µg/mL) aggregation of human washed platelets by SNALB (range, 0–10 µM) by cGMP-dependent and cGMP-independent mechanisms. The LMM thiols themselves did not affect platelet aggregation. It is assumed that the underlying mechanism involves S-transnitrosylation of SH groups of the platelet surface by LMM RSNO formed through the reaction of SNALB with the thiols: ALB-Cys34SNO + R-CysSH ↔ ALB-Cys34SH + R-CysSNO. Such S-transnitrosylation reactions may be accompanied by release of NO finally resulting in cGMP-dependent and cGMP-independent mechanisms.

Abstract Abstract 1095 Poster Board I-117 Introduction Fanconi anemia (FA) is a rare genetic disorder characterized by bone marrow failure, congenital abnormalities, and an increased risk for cancer and leukemia. Components of the FA-BRCA pathway are thought to function in the repair of DNA interstrand crosslinks (ICLs). Central to this pathway is the monoubiquitylation and chromatin localization of two FA proteins, FANCD2 and FANCI. Recent reports implicate mismatch repair factors in the repair of ICLs and have shown that FANCJ interacts with the MutLαa complex. Here we show that FANCD2 binds several mismatch repair proteins in vivo and that MSH2 is required for the monoubiquitylation and chromatin localization of both FANCD2 and FANCI. Methods Cell lines used: HeLa, human lung carcinoma cell line H1299, FA-A cell line GM6914 and corrected cell line GM6914 + Flag-FANCA, FA-D2 cell line PD20 and corrected cell lines PD20+Flag-FANCD2 and PD20+FANCD2 K561R, human endometrial adenocarcinoma cell line HEC59 (MSH2-deficient) and corrected cell line HEC59+Ch2, and human colon carcinoma cell line HCT116 (MLH1-deficient) and corrected cell line HCT116+Ch3. Cells were treated with the crosslinking agent mitomycin C (MMC). Immunoprecipitation was used to demonstrate the interaction between FANCD2 and MSH2, MLH1, and MSH3. Survival assays were performed by crystal violet staining and extraction. Chromatin loading of FANCD2 and FANCI was determined by cellular fractionation and western blot. Results Through chromatographic purification of FANCD2-containing protein complexes, we identified MSH2 and MLH1 as FANCD2-interacting proteins. Immunoprecipitation using HeLa cell extracts confirmed the interaction between FANCD2, MSH2, MSH3, and MLH1 in vivo. These interactions are all induced upon damage with a DNA crosslinking agent and MSH2 specifically interacts only with the monoubiquitylated form of FANCD2. Additionally, the FANCD2-MSH2 interaction requires ATR, but not ATM, BRCA1, MSH3, or ERCC1/XPF. Human cells lacking MSH2 display increased sensitivity to mitomycin C as compared to their corrected counterparts. FANCD2 and FANCI monoubiquitylation is also greatly diminished in these cells, while cells lacking MLH1 show no effect. Cellular fractionation of MSH2-deficient cells shows that FANCD2 and FANCI are not efficiently loaded onto chromatin after treatment with DNA-damaging agents, while MLH1-deficient cells again show no effect. Interestingly, while knockdown of either MSH2 or FANCD2 in H1299 cells results in increased sensitivity to MMC, double knockdown of both proteins corrects this sensitivity on par with controls. oConclusions These data suggest that mismatch repair proteins play a key role in the activation of the FA-BRCA pathway, likely through recognition of the DNA lesion. Understanding this role could lead to the development of new therapies for the treatment of patients both with FA and cancer. Disclosures No relevant conflicts of interest to declare.

Environments containing significant concentration of NaCl such as salt lakes harbor extremophiles microorganisms which have a great biotechnology interest. To explore the diversity of Bacteria in Chott Tinsilt (Algeria), an isolation program was performed. Water samples were collected from the saltern during the pre-salt harvesting phase. This Chott is high in salt (22.47% (w/v). Seven halophiles Bacteria were selected for further characterization. The isolated strains were able to grow optimally in media with 10–25% (w/v) total salts. Molecular identification of the isolates was performed by sequencing the 16S rRNA gene. It showed that these cultured isolates included members belonging to the Halomonas, Staphylococcus, Salinivibrio, Planococcus and Halobacillus genera with less than 98% of similarity with their closest phylogenetic relative. The halophilic bacterial isolates were also characterized for the production of biosurfactant and industrially important enzymes. Most isolates produced hydrolases and biosurfactants at high salt concentration. In fact, this is the first report on bacterial strains (A4 and B4) which were a good biosurfactant and coagulase producer at 20% and 25% ((w/v)) NaCl. In addition, the biosurfactant produced by the strain B4 at high salinity (25%) was also stable at high temperature (30-100°C) and high alkalinity (pH 11).Key word: Salt Lake, Bacteria, biosurfactant, Chott, halophiles, hydrolases, 16S rRNAINTRODUCTIONSaline lakes cover approximately 10% of the Earth’s surface area. The microbial populations of many hypersaline environments have already been studied in different geographical regions such as Great Salt Lake (USA), Dead Sea (Israel), Wadi Natrun Lake (Egypt), Lake Magadi (Kenya), Soda Lake (Antarctica) and Big Soda Lake and Mono Lake (California). Hypersaline regions differ from each other in terms of geographical location, salt concentration and chemical composition, which determine the nature of inhabitant microorganisms (Gupta et al., 2015). Then low taxonomic diversity is common to all these saline environments (Oren et al., 1993). Halophiles are found in nearly all major microbial clades, including prokaryotic (Bacteria and Archaea) and eukaryotic forms (DasSarma and Arora, 2001). They are classified as slight halophiles when they grow optimally at 0.2–0.85 M (2–5%) NaCl, as moderate halophiles when they grow at 0.85–3.4 M (5–20%) NaCl, and as extreme halophiles when they grow at 3.4–5.1 M (20–30%) NaCl. Hyper saline environments are inhabited by extremely halophilic and halotolerant microorganisms such as Halobacillus sp, Halobacterium sp., Haloarcula sp., Salinibacter ruber , Haloferax sp and Bacillus spp. (Solomon and Viswalingam, 2013). There is a tremendous demand for halophilic bacteria due to their biotechnological importance as sources of halophilic enzymes. Enzymes derived from halophiles are endowed with unique structural features and catalytic power to sustain the metabolic and physiological processes under high salt conditions. Some of these enzymes have been reported to be active and stable under more than one extreme condition (Karan and Khare, 2010). Applications are being considered in a range of industries such as food processing, washing, biosynthetic processes and environmental bioremediation. Halophilic proteases are widely used in the detergent and food industries (DasSarma and Arora, 2001). However, esterases and lipases have also been useful in laundry detergents for the removal of oil stains and are widely used as biocatalysts because of their ability to produce pure compounds. Likewise, amylases are used industrially in the first step of the production of high fructose corn syrup (hydrolysis of corn starch). They are also used in the textile industry in the de-sizing process and added to laundry detergents. Furthermore, for the environmental applications, the use of halophiles for bioremediation and biodegradation of various materials from industrial effluents to soil contaminants and accidental spills are being widely explored. In addition to enzymes, halophilic / halotolerants microorganisms living in saline environments, offer another potential applications in various fields of biotechnology like the production of biosurfactant. Biosurfactants are amphiphilic compounds synthesized from plants and microorganisms. They reduce surface tension and interfacial tension between individual molecules at the surface and interface respectively (Akbari et al., 2018). Comparing to the chemical surfactant, biosurfactant are promising alternative molecules due to their low toxicity, high biodegradability, environmental capability, mild production conditions, lower critical micelle concentration, higher selectivity, availability of resources and ability to function in wide ranges of pH, temperature and salinity (Rocha et al., 1992). They are used in various industries which include pharmaceuticals, petroleum, food, detergents, cosmetics, paints, paper products and water treatment (Akbari et al., 2018). The search for biosurfactants in extremophiles is particularly promising since these biomolecules can adapt and be stable in the harsh environments in which they are to be applied in biotechnology.OBJECTIVESEastern Algeria features numerous ecosystems including hypersaline environments, which are an important source of salt for food. The microbial diversity in Chott Tinsilt, a shallow Salt Lake with more than 200g/L salt concentration and a superficies of 2.154 Ha, has never yet been studied. The purpose of this research was to chemically analyse water samples collected from the Chott, isolate novel extremely or moderate halophilic Bacteria, and examine their phenotypic and phylogenetic characteristics with a view to screening for biosurfactants and enzymes of industrial interest.MATERIALS AND METHODSStudy area: The area is at 5 km of the Commune of Souk-Naâmane and 17 km in the South of the town of Aïn-Melila. This area skirts the trunk road 3 serving Constantine and Batna and the railway Constantine-Biskra. It is part the administrative jurisdiction of the Wilaya of Oum El Bouaghi. The Chott belongs to the wetlands of the High Plains of Constantine with a depth varying rather regularly without never exceeding 0.5 meter. Its length extends on 4 km with a width of 2.5 km (figure 1).Water samples and physico-chemical analysis: In February 2013, water samples were collected from various places at the Chott Tinsilt using Global Positioning System (GPS) coordinates of 35°53’14” N lat. and 06°28’44”E long. Samples were collected randomly in sterile polythene bags and transported immediately to the laboratory for isolation of halophilic microorganisms. All samples were treated within 24 h after collection. Temperature, pH and salinity were measured in situ using a multi-parameter probe (Hanna Instruments, Smithfield, RI, USA). The analytical methods used in this study to measure ions concentration (Ca2+, Mg2+, Fe2+, Na+, K+, Cl−, HCO3−, SO42−) were based on 4500-S-2 F standard methods described elsewhere (Association et al., 1920).Isolation of halophilic bacteria from water sample: The media (M1) used in the present study contain (g/L): 2.0 g of KCl, 100.0/200.0 g of NaCl, 1.0 g of MgSO4.7HO2, 3.0 g of Sodium Citrate, 0.36 g of MnCl2, 10.0 g of yeast extract and 15.0 g agar. The pH was adjusted to 8.0. Different dilutions of water samples were added to the above medium and incubated at 30°C during 2–7 days or more depending on growth. Appearance and growth of halophilic bacteria were monitored regularly. The growth was diluted 10 times and plated on complete medium agar (g/L): glucose 10.0; peptone 5.0; yeast extract 5.0; KH2PO4 5.0; agar 30.0; and NaCl 100.0/200.0. Resultant colonies were purified by repeated streaking on complete media agar. The pure cultures were preserved in 20% glycerol vials and stored at −80°C for long-term preservation.Biochemical characterisation of halophilic bacterial isolates: Bacterial isolates were studied for Gram’s reaction, cell morphology and pigmentation. Enzymatic assays (catalase, oxidase, nitrate reductase and urease), and assays for fermentation of lactose and mannitol were done as described by Smibert (1994).Optimization of growth conditions: Temperature, pH, and salt concentration were optimized for the growth of halophilic bacterial isolates. These growth parameters were studied quantitatively by growing the bacterial isolates in M1 medium with shaking at 200 rpm and measuring the cell density at 600 nm after 8 days of incubation. To study the effect of NaCl on the growth, bacterial isolates were inoculated on M1 medium supplemented with different concentration of NaCl: 1%-35% (w/v). The effect of pH on the growth of halophilic bacterial strains was studied by inoculating isolates on above described growth media containing NaCl and adjusted to acidic pH of 5 and 6 by using 1N HCl and alkaline pH of 8, 9, 10, 11 and 12 using 5N NaOH. The effect of temperature was studied by culturing the bacterial isolates in M1 medium at different temperatures of incubation (4°C–55°C).Screening of halophilic bacteria for hydrolytic enzymes: Hydrolase producing bacteria among the isolates were screened by plate assay on starch, tributyrin, gelatin and DNA agar plates respectively for amylase, lipase, protease and DNAse activities. Amylolytic activity of the cultures was screened on starch nutrient agar plates containing g/L: starch 10.0; peptone 5.0; yeast extract 3.0; agar 30.0; NaCl 100.0/250.0. The pH was 7.0. After incubation at 30 ºC for 7 days, the zone of clearance was determined by flooding the plates with iodine solution. The potential amylase producers were selected based on ratio of zone of clearance diameter to colony diameter. Lipase activity of the cultures was screened on tributyrin nutrient agar plates containing 1% (v/v) of tributyrin. Isolates that showed clear zones of tributyrin hydrolysis were identified as lipase producing bacteria. Proteolytic activity of the isolates was similarly screened on gelatin nutrient agar plates containing 10.0 g/L of gelatin. The isolates showing zones of gelatin clearance upon treatment with acidic mercuric chloride were selected and designated as protease producing bacteria. The presence of DNAse activity on plates was determined on DNAse test agar (BBL) containing 10%-25% (w/v) total salt. After incubation for 7days, the plates were flooded with 1N HCl solution. Clear halos around the colonies indicated DNAse activity (Jeffries et al., 1957).Milk clotting activity (coagulase activity) of the isolates was also determined following the procedure described (Berridge, 1952). Skim milk powder was reconstituted in 10 mM aqueous CaCl2 (pH 6.5) to a final concentration of 0.12 kg/L. Enzyme extracts were added at a rate of 0.1 mL per mL of milk. The coagulation point was determined by manual rotating of the test tube periodically, at short time intervals, and checking for visible clot formation.Screening of halophilic bacteria for biosurfactant production. Oil spread Assay: The Petridis base was filled with 50 mL of distilled water. On the water surface, 20μL of diesel and 10μl of culture were added respectively. The culture was introduced at different spots on the diesel, which is coated on the water surface. The occurrence of a clear zone was an indicator of positive result (Morikawa et al., 2000). The diameter of the oil expelling circles was measured by slide caliber (with a degree of accuracy of 0.02 mm).Surface tension and emulsification index (E24): Isolates were cultivated at 30 °C for 7 days on the enrichment medium containing 10-25% NaCl and diesel oil as the sole carbon source. The medium was centrifuged (7000 rpm for 20 min) and the surface tension of the cell-free culture broth was measured with a TS90000 surface tensiometer (Nima, Coventry, England) as a qualitative indicator of biosurfactant production. The culture broth was collected with a Pasteur pipette to remove the non-emulsified hydrocarbons. The emulsifying capacity was evaluated by an emulsification index (E24). The E24 of culture samples was determined by adding 2 mL of diesel oil to the same amount of culture, mixed for 2 min with a vortex, and allowed to stand for 24 h. E24 index is defined as the percentage of height of emulsified layer (mm) divided by the total height of the liquid column (mm).Biosurfactant stability studies : After growth on diesel oil as sole source of carbone, cultures supernatant obtained after centrifugation at 6,000 rpm for 15 min were considered as the source of crude biosurfactant. Its stability was determined by subjecting the culture supernatant to various temperature ranges (30, 40, 50, 60, 70, 80 and 100 °C) for 30 min then cooled to room temperature. Similarly, the effect of different pH (2–11) on the activity of the biosurfactant was tested. The activity of the biosurfactant was investigated by measuring the emulsification index (El-Sersy, 2012).Molecular identification of potential strains. DNA extraction and PCR amplification of 16S rDNA: Total cellular DNA was extracted from strains and purified as described by Sambrook et al. (1989). DNA was purified using Geneclean® Turbo (Q-BIO gene, Carlsbad, CA, USA) before use as a template in polymerase chain reaction (PCR) amplification. For the 16S rDNA gene sequence, the purified DNA was amplified using a universal primer set, forward primer (27f; 5′-AGA GTT TGA TCM TGG CTC AG) and a reverse primer (1492r; 5′-TAC GGY TAC CTT GTT ACG ACT T) (Lane, 1991). Agarose gel electrophoresis confirmed the amplification product as a 1400-bp DNA fragment.16S rDNA sequencing and Phylogenic analysis: Amplicons generated using primer pair 27f-1492r was sequenced using an automatic sequencer system at Macrogene Company (Seoul, Korea). The sequences were compared with those of the NCBI BLAST GenBank nucleotide sequence databases. Phylogenetic trees were constructed by the neighbor-joining method using MEGA version 5.05 software (Tamura et al., 2011). Bootstrap resembling analysis for 1,000 replicates was performed to estimate the confidence of tree topologies.Nucleotide sequence accession numbers: The nucleotide sequences reported in this work have been deposited in the EMBL Nucleotide Sequence Database. The accession numbers are represented in table 5.Statistics: All experiments were conducted in triplicates. Results were evaluated for statistical significance using ANOVA.RESULTSPhysico-chemical parameters of the collected water samples: The physicochemical properties of the collected water samples are reported in table 1. At the time of sampling, the temperature was 10.6°C and pH 7.89. The salinity of the sample, as determined in situ, was 224.70 g/L (22,47% (w/v)). Chemical analysis of water sample indicated that Na +and Cl- were the most abundant ions (table 1). SO4-2 and Mg+2 was present in much smaller amounts compared to Na +and Cl- concentration. Low levels of calcium, potassium and bicarbonate were also detected, often at less than 1 g/L.Characterization of isolates. Morphological and biochemical characteristic feature of halophilic bacterial isolates: Among 52 strains isolated from water of Chott Tinsilt, seven distinct bacteria (A1, A2, A3, A4, B1, B4 and B5) were chosen for further characterization (table 2). The colour of the isolates varied from beige, pale yellow, yellowish and orange. The bacterial isolates A1, A2, A4, B1 and B5 were rod shaped and gram negative (except B5), whereas A3 and B4 were cocci and gram positive. All strains were oxidase and catalase positive except for B1. Nitrate reductase and urease activities were observed in all the bacterial isolates, except B4. All the bacterial isolates were negative for H2S formation. B5 was the only strain positive for mannitol fermentation (table 2).We isolated halophilic bacteria on growth medium with NaCl supplementation at pH 7 and temperature of 30°C. We studied the effect of NaCl, temperature and pH on the growth of bacterial isolates. All the isolates exhibited growth only in the presence of NaCl indicating that these strains are halophilic. The optimum growth of isolates A3 and B1 was observed in the presence of 10% NaCl, whereas it was 15% NaCl for A1, A2 and B5. A4 and B4 showed optimum growth in the presence of 20% and 25% NaCl respectively. A4, B4 and B5 strains can tolerate up to 35% NaCl.The isolate B1 showed growth in medium supplemented with 10% NaCl and pH range of 7–10. The optimum pH for the growth B1 was 9 and they did not show any detectable growth at or below pH 6 (table 2), which indicates the alkaliphilic nature of B1 isolate. The bacterial isolates A1, A2 and A4 exhibited growth in the range of pH 6–10, while A3 and B4 did not show any growth at pH greater than 8. The optimum pH for growth of all strains (except B1) was pH 7.0 (table 2). These results indicate that A1, A2, A3, A4, B4 and B5 are neutrophilic in nature. All the bacterial isolates exhibited optimal growth at 30°C and no detectable growth at 55°C. Also, detectable growth of isolates A1, A2 and A4 was observed at 4°C. However, none of the bacterial strains could grow below 4°C and above 50°C (table 2).Screening of the halophilic enzymes: To characterize the diversity of halophiles able to produce hydrolytic enzymes among the population of microorganisms inhabiting the hypersaline habitats of East Algeria (Chott Tinsilt), a screening was performed. As described in Materials and Methods, samples were plated on solid media containing 10%-25% (w/v) of total salts and different substrates for the detection of amylase, protease, lipase and DNAse activities. However, coagulase activity was determined in liquid medium using milk as substrate (figure 3). Distributions of hydrolytic activity among the isolates are summarized in table 4.From the seven bacterial isolates, four strains A1, A2, A4 and B5 showed combined hydrolytic activities. They were positive for gelatinase, lipase and coagulase. A3 strain showed gelatinase and lipase activities. DNAse activities were detected with A1, A4, B1 and B5 isolates. B4 presented lipase and coagulase activity. Surprisingly, no amylase activity was detected among all the isolates.Screening for biosurfactant producing isolates: Oil spread assay: The results showed that all the strains could produce notable (>4 cm diameter) oil expelling circles (ranging from 4.11 cm to 4.67 cm). The average diameter for strain B5 was 4.67 cm, significantly (P < 0.05) higher than for the other strains.Surface tension and emulsification index (E24): The assimilation of hydrocarbons as the sole sources of carbon by the isolate strains led to the production of biosurfactants indicated by the emulsification index and the lowering of the surface tension of cell-free supernatant. Based on rapid growth on media containing diesel oil as sole carbon source, the seven isolates were tested for biosurfactant production and emulsification activity. The obtained values of the surface tension measurements as well as the emulsification index (E24) are shown in table 3. The highest reduction of surface tension was achieved with B5 and A3 isolates with values of 25.3 mN m−1 and 28.1 mN m−1 respectively. The emulsifying capacity evaluated by the E24 emulsification index was highest in the culture of isolate B4 (78%), B5 (77%) and A3 (76%) as shown in table 3 and figure 2. These emulsions were stable even after 4 months. The bacteria with emulsification indices higher than 50 % and/or reduction in the surface tension (under 30 mN/m) have been defined as potential biosurfactant producers. Based on surface tension and the E24 index results, isolates B5, B4, A3 and A4 are the best candidates for biosurfactant production. It is important to note that, strains B4 and A4 produce biosurfactant in medium containing respectively 25% and 20% (w/v) NaCl.Stability of biosurfactant activities: The applicability of biosurfactants in several biotechnological fields depends on their stability at different environmental conditions (temperatures, pH and NaCl). For this study, the strain B4 appear very interesting (It can produce biosurfactant at 25 % NaCl) and was choosen for futher analysis for biosurfactant stability. The effects of temperature and pH on the biosurfactant production by the strain B4 are shown in figure 4.biosurfactant in medium containing respectively 25% and 20% (w/v) NaCl.Stability of biosurfactant activities: The applicability of biosurfactants in several biotechnological fields depends on their stability at different environmental conditions (temperatures, pH and NaCl). For this study, the strain B4 appear very interesting (It can produce biosurfactant at 25 % NaCl) and was chosen for further analysis for biosurfactant stability. The effects of temperature and pH on the biosurfactant production by the strain B4 are shown in figure 4. The biosurfactant produced by this strain was shown to be thermostable giving an E-24 Index value greater than 78% (figure 4A). Heating of the biosurfactant to 100 °C caused no significant effect on the biosurfactant performance. Therefore, the surface activity of the crude biosurfactant supernatant remained relatively stable to pH changes between pH 6 and 11. At pH 11, the value of E24 showed almost 76% activity, whereas below pH 6 the activity was decreased up to 40% (figure 4A). The decreases of the emulsification activity by decreasing the pH value from basic to an acidic region; may be due to partial precipitation of the biosurfactant. This result indicated that biosurfactant produced by strain B4 show higher stability at alkaline than in acidic conditions.Molecular identification and phylogenies of potential isolates: To identify halophilic bacterial isolates, the 16S rDNA gene was amplified using gene-specific primers. A PCR product of ≈ 1.3 kb was detected in all the seven isolates. The 16S rDNA amplicons of each bacterial isolate was sequenced on both strands using 27F and 1492R primers. The complete nucleotide sequence of 1336,1374, 1377,1313, 1305,1308 and 1273 bp sequences were obtained from A1, A2, A3, A4, B1, B4 and B5 isolates respectively, and subjected to BLAST analysis. The 16S rDNA sequence analysis showed that the isolated strains belong to the genera Halomonas, Staphylococcus, Salinivibrio, Planococcus and Halobacillus as shown in table 5. The halophilic isolates A2 and A4 showed 97% similarity with the Halomonas variabilis strain GSP3 (accession no. AY505527) and the Halomonas sp. M59 (accession no. AM229319), respectively. As for A1, it showed 96% similarity with the Halomonas venusta strain GSP24 (accession no. AY553074). B1 and B4 showed for their part 96% similarity with the Salinivibrio costicola subsp. alcaliphilus strain 18AG DSM4743 (accession no. NR_042255) and the Planococcus citreus (accession no. JX122551), respectively. The bacterial isolate B5 showed 98% sequence similarity with the Halobacillus trueperi (accession no. HG931926), As for A3, it showed only 95% similarity with the Staphylococcus arlettae (accession no. KR047785). The 16S rDNA nucleotide sequences of all the seven halophilic bacterial strains have been submitted to the NCBI GenBank database under the accession number presented in table 5. The phylogenetic association of the isolates is shown in figure 5.DICUSSIONThe physicochemical properties of the collected water samples indicated that this water was relatively neutral (pH 7.89) similar to the Dead Sea and the Great Salt Lake (USA) and in contrast to the more basic lakes such as Lake Wadi Natrun (Egypt) (pH 11) and El Golea Salt Lake (Algeria) (pH 9). The salinity of the sample was 224.70 g/L (22,47% (w/v). This range of salinity (20-30%) for Chott Tinsilt is comparable to a number of well characterized hypersaline ecosystems including both natural and man-made habitats, such as the Great Salt Lake (USA) and solar salterns of Puerto Rico. Thus, Chott Tinsilt is a hypersaline environment, i.e. environments with salt concentrations well above that of seawater. Chemical analysis of water sample indicated that Na +and Cl- were the most abundant ions, as in most hypersaline ecosystems (with some exceptions such as the Dead Sea). These chemical water characteristics were consistent with the previously reported data in other hypersaline ecosystems (DasSarma and Arora, 2001; Oren, 2002; Hacěne et al., 2004). Among 52 strains isolated from this Chott, seven distinct bacteria (A1, A2, A3, A4, B1, B4 and B5) were chosen for phenotypique, genotypique and phylogenetique characterization.The 16S rDNA sequence analysis showed that the isolated strains belong to the genera Halomonas, Staphylococcus, Salinivibrio, Planococcus and Halobacillus. Genera obtained in the present study are commonly occurring in various saline habitats across the globe. Staphylococci have the ability to grow in a wide range of salt concentrations (Graham and Wilkinson, 1992; Morikawa et al., 2009; Roohi et al., 2014). For example, in Pakistan, Staphylococcus strains were isolated from various salt samples during the study conducted by Roohi et al. (2014) and these results agreed with previous reports. Halomonas, halophilic and/or halotolerant Gram-negative bacteria are typically found in saline environments (Kim et al., 2013). The presence of Planococcus and Halobacillus has been reported in studies about hypersaline lakes; like La Sal del Rey (USA) (Phillips et al., 2012) and Great Salt Lake (Spring et al., 1996), respectively. The Salinivibrio costicola was a representative model for studies on osmoregulatory and other physiological mechanisms of moderately halophilic bacteria (Oren, 2006).However, it is interesting to note that all strains shared less than 98.7% identity (the usual species cut-off proposed by Yarza et al. (2014) with their closest phylogenetic relative, suggesting that they could be considered as new species. Phenotypic, genetic and phylogenetic analyses have been suggested for the complete identification of these strains. Theses bacterial strains were tested for the production of industrially important enzymes (Amylase, protease, lipase, DNAse and coagulase). These isolates are good candidates as sources of novel enzymes with biotechnological potential as they can be used in different industrial processes at high salt concentration (up to 25% NaCl for B4). Prominent amylase, lipase, protease and DNAase activities have been reported from different hypersaline environments across the globe; e.g., Spain (Sánchez‐Porro et al., 2003), Iran (Rohban et al., 2009), Tunisia (Baati et al., 2010) and India (Gupta et al., 2016). However, to the best of our knowledge, the coagulase activity has never been detected in extreme halophilic bacteria. Isolation and characterization of crude enzymes (especially coagulase) to investigate their properties and stability are in progress.The finding of novel enzymes with optimal activities at various ranges of salt concentrations is of great importance. Besides being intrinsically stable and active at high salt concentrations, halophilic and halotolerant enzymes offer great opportunities in biotechnological applications, such as environmental bioremediation (marine, oilfiel) and food processing. The bacterial isolates were also characterized for production of biosurfactants by oil-spread assay, measurement of surface tension and emulsification index (E24). There are few reports on biosurfactant producers in hypersaline environments and in recent years, there has been a greater increase in interest and importance in halophilic bacteria for biomolecules (Donio et al., 2013; Sarafin et al., 2014). Halophiles, which have a unique lipid composition, may have an important role to play as surface-active agents. The archae bacterial ether-linked phytanyl membrane lipid of the extremely halophilic bacteria has been shown to have surfactant properties (Post and Collins, 1982). Yakimov et al. (1995) reported the production of biosurfactant by a halotolerant Bacillus licheniformis strain BAS 50 which was able to produce a lipopeptide surfactant when cultured at salinities up to 13% NaCl. From solar salt, Halomonas sp. BS4 and Kocuria marina BS-15 were found to be able to produce biosurfactant when cultured at salinities of 8% and 10% NaCl respectively (Donio et al., 2013; Sarafin et al., 2014). In the present work, strains B4 and A4 produce biosurfactant in medium containing respectively 25% and 20% NaCl. To our knowledge, this is the first report on biosurfactant production by bacteria under such salt concentration. Biosurfactants have a wide variety of industrial and environmental applications (Akbari et al., 2018) but their applicability depends on their stability at different environmental conditions. The strain B4 which can produce biosurfactant at 25% NaCl showed good stability in alkaline pH and at a temperature range of 30°C-100°C. Due to the enormous utilization of biosurfactant in detergent manufacture the choice of alkaline biosurfactant is researched (Elazzazy et al., 2015). On the other hand, the interesting finding was the thermostability of the produced biosurfactant even after heat treatment (100°C for 30 min) which suggests the use of this biosurfactant in industries where heating is of a paramount importance (Khopade et al., 2012). To date, more attention has been focused on biosurfactant producing bacteria under extreme conditions for industrial and commercial usefulness. In fact, the biosurfactant produce by strain B4 have promising usefulness in pharmaceutical, cosmetics and food industries and for bioremediation in marine environment and Microbial enhanced oil recovery (MEOR) where the salinity, temperature and pH are high.CONCLUSIONThis is the first study on the culturable halophilic bacteria community inhabiting Chott Tinsilt in Eastern Algeria. Different genera of halotolerant bacteria with different phylogeneticaly characteristics have been isolated from this Chott. Culturing of bacteria and their molecular analysis provides an opportunity to have a wide range of cultured microorganisms from extreme habitats like hypersaline environments. Enzymes produced by halophilic bacteria show interesting properties like their ability to remain functional in extreme conditions, such as high temperatures, wide range of pH, and high salt concentrations. 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Introduction: Polycyclic aromatic hydrocarbons (PAHs) are prevalent organic pollutants that garner attention due to their toxicity and carcinogenic properties. These hydrophobic compounds tend to accumulate in the organic components of sediments, posing significant ecological risks. Methods: This study synthesizes analytical methodologies and assesses the contamination status of PAHs in Vietnamese sediments through a review of previous studies complemented by recent findings from our research group. Various extraction techniques, including Soxhlet extraction, pressurized liquid extraction, continuous solid-liquid extraction, and ultrasonic extraction, were employed using organic solvents of low to moderate polarity (e.g., hexane, acetone, toluene, dichloromethane). Purification was achieved via column chromatography using sorbents like silica gel, alumina, and Florisil, followed by gel permeation chromatography. Separation and analysis were primarily conducted through gas chromatography/mass spectrometry (GC/MS). Results: PAHs were detected in all samples, with concentrations ranging from 33 to 6400 ng/g, indicating widespread contamination. The sources of PAHs in Vietnamese sediments appear to be predominantly urban and informal waste processing areas. The levels of sedimentary PAHs in Vietnam are on par with those reported in other Asian countries, with the majority of PAHs being traced back to pyrogenic sources, as opposed to petroleum products. Conclusions: Our findings offer a detailed understanding of the distribution, patterns, and potential sources of PAHs in Vietnamese sediments. The notably higher concentrations of PAHs in urban and waste processing sites underscore the urgency of implementing effective management and remediation strategies. The study advocates for further monitoring of PAHs and related pollutants to fully elucidate their contamination scope, origins, and ecological consequences.

Abstract An appropriate purification and quantification method has been developed for polycyclic aromatic hydrocarbons (PAH) in hydro alcoholic herbal extracts. For this, Bacopa monnieri, Camellia sinensis, Withania somnifera and Andrographis paniculata samples were extracted with modified solid-phase extraction (SPE) and the PAH were quantified using liquid chromatography coupled to fluorescence detector. Purification of herbal extract using hexane and acetone in the ratio of 1:1 followed by treatment with QuEChERS salt (6 g MgSO4 and 1.5 g sodium acetate) improved the recovery rate of PAH. Silica SPE, which accomplishes solvent exchange to hexane by cleanup method, was developed to reduce the matrix effect and quality of the result obtained was increased. The developed method can be used for regular monitoring and analysis of PAH in natural extracts so as to prevent contamination.

Introduction : Les rayons ultraviolet (UVR) peuvent-être nocifs pour la peau, par l’action des radicaux libres générés par les espèces réactives de l’oxygène (ROS), comme l’oxygène singulet (1O2). Le beurre de karité (BK) revendique des propriétés de photoprotection contre les rayons UVA et UVB, par l’action de ses insaponifiables. Notre étude consiste à déterminer la capacité antioxydante du BK extrait au Burkina Faso, et à évaluer in vitro son pouvoir de photoprotection indirecte. Matériel et Méthodes : Les composés phénoliques totaux du BK brut, ont été obtenus par extraction liquide-liquide et dosés par technique colorimétrique avec le réactif de Folin-Ciocalteu. Leur capacité antioxydante a été évaluée par les tests colorimétriques ABTS et DPPH. Un modèle expérimental UVR- like a été mis au point afin d’évaluer in vitro le pouvoir photo-protecteur indirecte du beurre brut et de ses extraits phénoliques par photosensibilisation du Rose Bengal (RB) en présence de l’anthracène. Résultats : La teneur en phénols totaux des échantillons de BK du Burkina Faso était comprise entre 14,16 ± 2,33 et 82,99 ± 8,70 ppm d'équivalent pyrogallol. Ces composés phénoliques ont montré une activité antiradicalaire dose-dépendante. Sur modèle acellulaire expérimental UVR-like, le beurre brut et sa fraction phénolique ont montré une capacité quenching dose-dépendante de l’1O2 par photosensibilisation du RB, en présence d’anthracène, ce qui met en évidence une photoprotection indirecte du beurre brut et de ses composés phénoliques. Conclusion : Le BK brut peut être utilisé pour ses propriétés antioxydantes dû à sa fraction phénolique. Outre ses propriétés antioxydantes, le BK a un pouvoir de photoprotection indirecte, par effet quenching de l’O2, qui est plus prononcé que celui de sa fraction phénolique. L'effet photoprotecteur sur modèle UVR-like pourrait donc être attribuable à un effet antioxydant indirecte, et à l’action des insaponifiables.

AbstractOrange peels (OPs) were valorized in a lab-scale biorefinery loop for the recovery of limonene and the subsequent production of volatile fatty acids (VFAs) and activated carbon (AC). Solid/liquid extraction of limonene was optimized using n-hexane at 85 °C with an OPs-to-solvent ratio of 2:1, allowing for a limonene recovery yield of 1.20% w/w. Then, post-extraction OPs were used for the production of both VFAs and AC. For VFA production, a hydraulic retention time (HRT) of 5 days and a total solid (TS) inlet content of 10% w/w were adopted leading to a VFA yield of about 43% gVFAs/gTS. Adsorption tests revealed that, among all the solid matrixes tested, only powdered activated carbon (PAC) was able to discriminate no-VFA compounds and allowed for VFA purification. For AC production, post-extraction OPs were firstly converted into biochar through slow pyrolysis at 550 °C for 1 h and then physically activated with CO2 at 880 °C for 1 h. Extraction did not appreciably affect OP properties, while pyrolysis increased the carbon content (from 43 to 83%) and the heating value (from 17 to 29 MJ/kg) of the material. Physical activation of OP biochar increased its surface area by almost ten times, from 40 to 326 m2/g, proving the effectiveness of the treatment.

Introduction : La présente étude avait pour objectif d’évaluer l’effet sur le glucose sanguin d’un extrait hydro-éthanolique des écorces de F. albida (Fabacées) sur un modèle d’étude du diabète. Matériel et méthodes : La poudre des écorces de Faidherbia albida a été traitée par extraction hydro-éthanolique. L’extrait hydro-alcoolique (EHA) obtenu a été caractérisé au plan phytochimique et testé chez des rats normoglycémiques, sur un test de tolérance au glucose et chez des rats diabétiques de type 2. Résultats : L’EHA renfermant des flavonoïdes, des tanins et dépourvu d’alcaloïdes, est sans effet sur la glycémie de base des rats normoglycémiques (0,88 ± 0,10 vs 0,77 ± 0,15 g/l) et (0,87 ± 0,03 vs 0,74 ± 0,06 g/L) respectivement aux doses de 100mg/kg et 300mg/kg per os. En revanche, il est anti-hyperglycémiant de façon dépendante de la dose sur un test de tolérance au glucose. En effet, aux doses de 100 et 300 mg/kg per os de l’EHA, le pic hyperglycémique après administration de glucose (4 g/kg, per os) était respectivement 1,21 ± 0,05 g/L et 1,04 ± 0,10 g/L vs 2,03 ± 0,28 g/L dans le groupe de contrôle. L’EHA est anti-hyperglycémiant sur un modèle de troubles de la sécrétion d’insuline induit par l’alloxane. En effet, à la dose de 100mg/kg per os, le glucose sanguin, au bout de huit (08) jours d’observation, était de 2,38 ± 0,55 g/L vs 3,82 ± 0,52 dans le groupe témoin (eau physiologique) et 1,91 ± 1,25 celui du groupe traité par le glibenclamide. Conclusion : Les composés des écorces de Faidherbia albida sont exclusivement anti-hyperglycémiants, probablement par un mécanisme mettant en jeu une amélioration de l’action de l’insuline au niveau des tissus périphériques. L’effet anti-hyperglycémiant de l’EHA pourrait être lié à la présence de flavonoïdes dans l’extrait. En perspective, nous envisageons de poursuive le fractionnement de l’extrait alcoolique par chromatographie liquide haute performance afin d’isoler la molécule présentant un intérêt dans le traitement du diabète de type 2

This research aimed to recover anthocyanin-rich extracts from blackberry (Rubus spp. Hull cultivar) by optimizing the processing conditions, and to characterize anthocyanin individuals and determine influences of optimization on enhancement of antioxidant and anti-hyperglycemic activities of anthocyanins as natural supplements. The ethanol concentration of 69.87%, HCl dosage of 0.53%, solid-to-liquid ratio of 1:19.06 at 47.68°C for 17.04 h were optimal to obtain the highest extraction yield of anthocyanins at 0.72 mg/g. By using AB-8 macroporous resins, the anthocyanin concentration of 3.0 mg/mL, ethanol concentration of 90%, and elution rate of 2.0 mL/min were selected to boost the anthocyanin purity up to be 60.11%. Moreover, the purified anthocyanin extracts from blackberry contained nine main pigments which could be divided into three aglycone-based forms, and cyanidin-3-O-glucoside was the most abundant among them. Due to the successive processes of extraction and purification, the blackberry purified anthocyanin extracts (BA-PAE) showed much higher bioactive capacities than the blackberry crude anthocyanin extracts (BA-CAE) and blackberry fruit slurry extracts (BA-FSE), e.g., DPPH and ABTS radical scavenging activities (EC50 = 0.08 and 0.04, 0.32 and 0.24, and 1.31 and 0.41 mg/mL), oxygen radical absorbance capacity (1.60, 0.59, and 0.15 mmol TEAC/g), cytoprotective effects against oxidative stress in PC12 cells (1.69-, 1.58-, and 1.50-fold cell viability compared to oxidative group), α-amylase and α-glucosidase inhibitory activities (IC50 = 0.10 and 0.06, 0.56 and 0.32, and 3.98 and 2.16 mg/mL), and antibacterial activity (93.23, 40.85, and 80.42% reduced biofilm).

Abstract Background Resource-limited regions face a greater burden of infectious diseases due to limited access to molecular tests, complicating timely diagnosis and management. Current molecular point-of-care tests (POCTs) either come with high costs or lack adequate sensitivity and specificity. To facilitate better prevention and control of infectious diseases in underserved areas, we seek to address the need for molecular POCTs that better align with the World Health Organization (WHO)’s ASSURED criteria—Affordable, Sensitive, Specific, User-friendly, Rapid and robust, Equipment-free, and Deliverable to end users. Methods A novel molecular POCT, Pasteur Pipette-assisted isothermal probe amplification (pp-IPA), was developed for malaria detection. Without any microfluidics, this method captures Plasmodium 18S rRNA in a modified Pasteur pipette using tailed genus-specific probes. After washing, the bound tailed probes are ligated to form a template for subsequent novel isothermal probe amplification using a pair of generic primers, bypassing nucleic acid extraction and reverse transcription. The method was assessed using cultured Plasmodium and compared with real-time quantitative reverse transcription PCR (RT-qPCR) or reverse transcription loop-mediated isothermal amplification (RT-LAMP) in clinical blood samples. Results The entire assay is completed in 60–80 min with minimal hands-on time, using only a Pasteur pipette and a water bath. The pp-IPA’s analytical sensitivity is 1.28 × 10–4 parasites/μl, with 100% specificity against various blood-borne pathogens causing malaria-like symptoms. Additionally, pp-IPA needs only liquid-transfer skill for operation and the cost is around USD 0.25 per test, making it at least 300 times lower than mainstream POCT platforms. Conclusions Designed to improve the accessibility of molecular detection in resource-limited settings, pp-IPA’s simplicity, affordability, high sensitivity/specificity, and minimal equipment requirements make it a promising point-of-care pathogen identification tool in resource-constrained regions. Graphical Abstract